Characteristics of cytotoxicity of native zeolite against cattle epithelial cells

Abstract

Relevance. In recent years, there has been an increasing interest in the study of trepel, the mass fraction of zeolite in which reaches 50%. The purpose of this study was to assess the cytotoxic effect of the Zikeyevsky field trepel on the cells of the cow embryo lung (CEL).Methods. To characterize cytotoxicity, 6 fractions of trepel were selected (mining quarry — Zikeyevskoye deposit of Zhizdrinsky district of Kaluga region) with particle sizes: no. 1 — 0.03 mm, no. 2 — 0,03–0,06 mm, no. 3 — 0,09–0,15 mm, no. 4 — 0,15–0,3 mm, no. 5 — 0,3–0,5 mm. To determine cytotoxicity, zeolite suspensions were prepared on a DMEM maintenance medium in concentrations 50, 100, 200, 300, 400, 500, 1000 micrograms/ ml. As a model for determining the toxicity of trepeles, a transplanted cell line of CEL (epithelium of the lung embryo of a cow) obtained from a Collection of vertebrate cell cultures (ICiG RAS, Russia) was selected. The cells were grown on a DMEM medium with the addition of 10% fetal bovine serum ("HyClone", USA), penicillin and streptomycin (100 IU/ml each), 20 mM glutamine ("Sigma-Aldrich", USA) in 96-well tablets ("Costar", USA) at +37 °C in an atmosphere of 5% CO2. Cell viability was assessed by the activity of mitochondrial dehydrogenase according to the standard method. Statistical data processing was performed using "GraphPad Prism 6.0" software (USA). The experiments were carried out in 5 repeats, p ≤ 0.05 was taken as a statistically significant level.Results. It was found that concentrations of all fractions of trepels up to 100 micrograms/ml do not have a cytotoxic effect on the studied cells. A significant cytopathogenic effect is manifested in the concentration range of 300–1000 micrograms/ml; is in a positive correlation with the size of trepel particles — the proportion of necrotic cells when processing monolayers with maximum concentrations is up to 99.7±2.9%, which may be due to the mechanical effect of large mineral particles on cellular structures.